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Description
Human Sialin (AST) ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: Tissue homogenization: Rinse the tissue with pre chilled PBS (0. 01M, pH 7. 4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh and
Product Specification
| Usage | Required experimental equipment: 1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: Tissue homogenization: Rinse the tissue with pre-chilled PBS (0.01M, pH 7.4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL of PBS. The specific volume can be adjusted according to experimental needs and recorded. It is recommended to add protease inhibitors to the PBS) in a glass homogenizer and grind thoroughly on ice. To further lyse tissue cells, the homogenate can be sonicated or repeatedly frozen and thawed. Finally, centrifuge the homogenate at 5000×g for 5-10 minutes, and collect the supernatant for analysis. Cell Lysis Buffer: Adherent cells should be gently washed with pre-chilled PBS, then trypsinized and harvested by centrifugation at 1000×g for 5 minutes. Suspension cells can be harvested directly by centrifugation. Collected cells should be washed three times with pre-chilled PBS and resuspended in 150-200 μL of PBS per 1×10^6 cells (it is recommended to add protease inhibitors to the PBS; if the cell count is very low, reduce the PBS volume appropriately). Disrupt the cells by repeated freezing and thawing or sonication. Centrifuge the extract at 1500×g for 10 minutes at 2-8°C, and collect the supernatant for analysis. Other biological fluids: Centrifuge at 1000xg for 20 minutes, remove the supernatant, and test. Pre-test preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the standard gradient working solution: Add 1 mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 20 ng/mL). Then dilute to the following concentrations: 20 ng/mL, 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, and 0 ng/mL. Serial dilution method: Take 7 EP tubes and add 500 μL of universal diluent to each tube. Pipette 500 μL of the 20 ng/mL standard working solution into the first EP tube and mix thoroughly to make a 10 ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves directly as a blank well; there is no need to aspirate the liquid from the penultimate tube. See the figure below for details. 3. Preparation of Biotinylated Antibody Working Solution: 15 minutes before use, centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration using universal diluent (e.g., 10µL concentrate + 990µL universal diluent). Prepare immediately before use. 4. Prepare the enzyme conjugate working solution: 15 minutes before use, centrifuge the 100× concentrated enzyme conjugate at 1000×g for 1 minute. Dilute the 100× concentrated HRP enzyme conjugate to a 1× working concentration with universal diluent (e.g., 10 μL of concentrate + 990 μL of universal diluent). Prepare immediately. 5. Prepare the 1× wash solution: Dispense 10 mL of 20× wash solution into 190 mL of distilled water (concentrated wash solution removed from the refrigerator may crystallize; this is normal. Allow to stand at room temperature until the crystals have completely dissolved before preparing). Procedure: 1. Remove the desired strips from the aluminum foil bag after equilibration at room temperature for 10 minutes. Seal the remaining strips in a ziplock bag and return to 4°C. 2. Sample addition: Add 100 μL of sample or standard of varying concentrations to the corresponding wells. Add 100 μL of universal diluent to the blank wells. Cover with a film and incubate at 37°C for 60 minutes. (Recommendation: Dilute the sample to be tested at least 1-fold with universal diluent before adding it to the ELISA plate. This will reduce the impact of matrix effects on the test results. The sample concentration should be multiplied by the corresponding dilution factor when calculating the final sample concentration. It is recommended to run replicates for all test samples and standards.) 3. Add Biotinylated Antibody: Remove the ELISA plate and discard the liquid without washing. Add 100 μL of Biotinylated Antibody Working Solution directly to each well. Cover with a film and incubate at 37°C for 60 minutes. 4. Wash: Discard the liquid and add 300 μL of 1x Wash Solution to each well. Let stand for 1 minute, shake off the wash solution, and pat dry on absorbent paper. Repeat this process three times (a plate washer can also be used). 5. Add Enzyme Conjugate Working Solution: Add 100 μL of Enzyme Conjugate Working Solution to each well. Cover with a film and incubate at 37°C for 30 minutes. 6. Washing: Discard the liquid and wash the plate five times as in step 4. 7. Adding substrate: Add 90 μL of substrate (TMB) to each well, cover with a sealing film, and incubate at 37°C in the dark for 15 minutes. 8. Adding stop solution: Remove the ELISA plate and add 50 μL of stop solution directly to each well. Immediately measure the OD value of each well at a wavelength of 450 nm. Calculating experimental results: 1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as a correction factor. Plot the standard curve of the four-parameter logistic function on double-logarithmic graph paper, with concentration as the horizontal axis and OD value as the vertical axis. 2. If the sample OD value is higher than the upper limit of the standard curve, dilute the sample appropriately and retest. Multiply the sample concentration by the corresponding dilution factor. |
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| Sensitivity | 0.16 ng/mL | |||||||||||||||||||||||||||||||||
| Theory | This kit uses a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a sialin (AST) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB converts to blue under the catalysis of HRP and to yellow under the action of acid. The intensity of the color is positively correlated with the amount of sialin (AST) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Human | |||||||||||||||||||||||||||||||||
| Synonym | Human Sialin ELISA Kit; H(+)/nitrate cotransporter, H(+)/sialic acid cotransporter (AST), Membrane glycoprotein HP59, Solute carrier family 17 member 5 (SLC17A5), Vesicular excitatory amino acid transporter | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | The anion/sugar transporter (AST), also known as sialic acid or solute carrier family 17 member 5 (SLC17A5), is a protein encoded by the SLC17A5 gene. It cleaves glucuronic acid and free sialic acid from degrading sialic acid glycoconjugates and transports them out of lysosomes, which is essential for normal CNS myelination. It mediates the membrane potential-dependent uptake of aspartate and glutamate into synaptic vesicles and synaptic-like microvesicles. | |||||||||||||||||||||||||||||||||
| General Notes | 1. Strictly adhere to the specified incubation time and temperature to ensure accurate results. All reagents must be at room temperature (20-25°C) before use. Refrigerate reagents immediately after use. 2. Improper plate washing may result in inaccurate results. Ensure that all liquid in the wells is aspirated thoroughly before adding substrate. Do not allow the wells to dry out during incubation. 3. Remove any residual liquid and fingerprints from the bottom of the plate, as this will affect the OD value. 4. The substrate developer solution should be colorless or very light in color. Do not use substrate solution that has turned blue. 5. Avoid cross-contamination of reagents and specimens to prevent erroneous results. 6. Avoid direct exposure to strong light during storage and incubation. 7. Do not expose any reagents to bleaching solvents or the strong fumes emitted by bleaching solvents. Any bleaching agent will destroy the biological activity of the reagents in the kit. 8. Do not use expired products, and do not mix components with different product numbers and batches. 9. Recombinant proteins from sources other than the kit may not be compatible with the antibodies in this kit and will not be recognized. 10. If there is a possibility of disease transmission, all samples should be managed properly and samples and testing devices should be handled according to prescribed procedures. |
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| Storage Temp. | If the unopened kit is stored at 4°C, the shelf life is 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 0.312-20 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates, cell lysates, and other biological fluids |
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4.7 ★★★★★
Based on 1523 reviews
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Product Reviews
★★★★★ 3
Gothic Star Wars
Format: Paperback
This trade paperback collects all the issues for the Screaming Citadel story spread over several titles, including the main stay Star Wars series and the Dr Aphra book. As one might expect from a story spread over different titles with different artists and writers, the presentation varies. The art is all over the place. In the Marco Checchetto-drawn initial issue, everyone’s favorite amoral artifact hunter, Dr Aphra, is a striking space vixen. But in the following issues she’s hardly recognizable as the same character--mousier, if still menacing, in her trademark Russian tanker’s hat. To a lesser degree, the same is true for the other characters, including the main SW group. It’s understandable, but a bit disconcerting.
The story centers on Dr Aphra, who, in need of a Jedi for one of her typically nefarious purposes, recruits Luke into her scheme. Unfortunately for Aphra, she’s up against a more ruthless foe in the harlequin-looking vampire-like Queen of the Screaming Citadel. Before long, the rest of the group has to show up to rescue them. It’s a gothic story, set in scary castle—not the usual Star Wars fare. There are some good points. Dr Aphra’s almost sociopathic outlook is always good for a few choice lines, the “murderous machines” Bee Tee and Triple Zero are on hand for their own gruesome commentary and some of the Queens hench-people, while not given much to do, are interestingly designed. But overall, the horror movies plotline didn’t seem much like Star Wars to me. Recommended for those who enjoy that type of story, or completists.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on February 27, 2018
★★★★★ 4
Luke and Doctor Aphra team up!
Format: Paperback
This is the second crossover event in the Marvel Star Wars comics. It brings the ongoing Doctor Aphra and Star Wars series together. I liked the pairing of Luke with Aphra. They play well off of each other with Luke's naive goodness and Aphra's experienced gray morality. I liked when she called him a wannabe padawan.
There are some well designed characters in this comic. The residents of the Screaming Citadel have a goth bdsm vibe. Luke even gets to dress up. I liked seeing him in something different.
I want to know more about Sana and Aphra's past!!! Please, Marvel, make a queer love story prequel!!!
The murder droids are wonderful. Having them on the same side as the "good guys" for at least the time being led to some funny situations.
The last panel intrigued me. I give this graphic novel a 4/5. I am always here for more Doctor Aphra!
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Reviewed in the United States on December 29, 2017
★★★★★ 5
Excellent mini series.
Format: Kindle
This is an excellent follow up to Vader Down. Luke Skywalker and friends take on a bigger threat than The Empire and Darth Vader that is connected to the Jedi. Luke and Dr. Aphra join forces to find the answers Like is seeking. Truly worth reading and entertaining.
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on June 23, 2019
★★★★★ 5
Source material for Nolan's trilogy is powerful writing and art...
Format: Paperback
The Long Halloween is great, but I actually had more fun reading through Dark Victory. It's a crime mystery that consistently had me guessing through to the conclusion about who The Hangman's true identity was. What I really loved most was the evolution of the characters' personalities--from Jim Gordon to Harvey Dent to Bruce (who really is more in his iconic "Batman" persona here)--that began in Batman: Year One to Batman: The Long Halloween and into Batman: Dark Victory. It really does work like a trilogy. It's also notable to include Gotham city's underworld itself as a character of its own. Batman begins (pardon the pun) in Year One with a quest to sort of rid the city of the mafia, which is and has been the main criminal body up until this point. In The Long Halloween, the mafia begins to lose power because of the rise of the Batman's presence. During that time, some of the most insane and dangerous criminals escape Arkham Asylum and begin quietly terrorizing the city anew, spreading like a virus. Slowly, the "freaks"--as the mafia calls them--start to gain more and more power by simply being a more unorganized crime source (as opposed to organized crime). Characters like Pamela Isley, Solomon Grundy, Mr. Freeze, the Penguin, Scarecrow, the Joker, and others, establish and strengthen their grips on Gotham's criminal underworld.
It's nice that these villains--these "freaks"--also aren't the main conflict in Dark Victory (or The Long Halloween or Year One, for that matter); they appear when it is effective for them to appear, slowly taking more prominence in the setting of the story as it progresses. The main conflict has to deal with the solving of the mysteries behind The Hangman killings. The Long Halloween and Dark Victory are, primarily, crime mysteries, which is what makes them interesting. What makes them great literature, however, is the creative team that is Jeph Loeb and Tim Sale.
Loeb's writing is really well-crafted here: the entire arc unfolds at a consistent pace and he balances out dialogue with Batman's internal narration very smoothly. And, as I mentioned previously, personality and narrative arcs of the main characters (Gordon, Batman, and Dent) have fully matured by this point in the trilogy, leaving the supporting characters a chance to evolve and come into their own.
Much of these progressions are not just depicted by the writing, however; Tim Sale (who worked previously with Loeb on The Long Halloween) elevates and perfects his artwork in this story. There's not a change in the look of the characters, so you know it's definitely his style, but you sense immediately (especially if reading Dark Victory right after finishing The Long Halloween) the new level of attention paid to composition of the drawings. Shadows and silhouettes, contrasts between setting and characters, everything adds to the mood and atmosphere of the characters and the scenes they're a part of.
In terms of the print itself, the paperback is excellent. This and The Long Halloween have really nice paper, which I'll catch myself sometimes randomly sniffing in the middle of a read to enjoy the new paper smell. The ink is really crisp, the colors pop, and the design of the book itself is laid out very clearly. There aren't page numbers or a contents page, but every issue is separated by chapter pages that include gorgeous, high-contrast artwork to help distinguish which issue you're on. Additionally, the print comes with an introduction by David S. Goyer, who co-wrote the Nolan film trilogy.
Overall, I can't stress enough how gorgeous this trade paperback is and how excellent and top-tier this story arc is. I really enjoyed it just as much, if not more so, then The Long Halloween. No other Batman story arc has topped my enjoyment of this particular trilogy. Highly recommended in addition to Batman: Year One and Batman: The Long Halloween.
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Reviewed in the United States on June 9, 2019
★★★★★ 5
I love this comic book!
Format: Paperback, Format: Paperback
I love DC comics and obviously the Batfam are some of the most well known and loved characters within the DC universe. I love the art style and story in this comic. If you are debating whether or not to purchase this comic, DO IT!
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on December 23, 2025
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